Single nucleotide polymorphism detection in promoter I of the acetyl-CoA carboxylase-α gene in sheep

During analysis of the genome of four breeds of sheep - Gentile di Puglia, Sopravissana, Sarda and Romanoff - three single nucleotide polymorphisms (SNPs) were detected in the 5′ untranslated region of locus OAR292285 encoding the DNA sequence of promoter I of the acetyl-CoA carboxylase-α gene. DNA analysis was performed through denaturing high-performance liquid chromatography (DHPLC) using the WAVE® Nucleic Acid Fragment Analysis System. All SNPs were subsequently confirmed by direct sequencing. We demonstrated that the DNA profiles detected through DHPLC allowed discrimination between different genotypes even when multiple SNPs were found within the same amplicon. Allele frequency at each detected SNP was calculated. None of the detected SNPs allowed discrimination between breeds, because at least one individual of each analysed breed possessed each detected allele. However, the Sopravissana breed showed significantly different allele frequencies from the Sarda and the Gentile di Puglia breeds.